Method for producing transfructosylated steviol glycoside using lactobacillus mali
A technology of steviol and glycosides, which is applied in the field of preparing transglucosylated steviol glycosides, can solve the problems of increasing calories and the like, and achieve the effect of effective production
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Embodiment 1
[0046] Example 1: Preparation method of a novel enzyme derived from Lactobacillus malipus with sugar hydrolysis activity
[0047] In a nutrient medium containing yeast extract and corn steep liquor as a nitrogen source, sugar (Beksul white sugar with a purity of 99% or more) as a carbon source, amino acids, etc., the Lactobacillus malus microorganism (DSM20444 , ATCC27054, ATCC 27304) 24 hours. The medium (culture) was centrifuged at 8000 rpm for 10 minutes to separate cells and supernatant, and then only the supernatant was collected. The crude enzyme liquid was reacted with sugar, which resulted in sugar hydrolyzing activity, and this confirmed the presence of a novel enzyme having sugar hydrolyzing activity in the crude enzyme liquid.
Embodiment 2
[0048] Example 2: Evaluation of the conversion from steviol glycosides to transglucosylated steviol glycosides
[0049] Steviol glycosides and sugars were dissolved in a 0.05M acetate buffer solution, and the crude enzyme solution of the Lactobacillus malica strain prepared in Example 1 was added thereto, followed by reaction at 40° C. for 24 hours. After the reaction, the reaction solution was inactivated at 100° C., and then production of transglucosylated steviol glycoside was confirmed by HPLC. The steviol glycosides used were stevioside, rubusoside, dulcoside A, and rebaudioside A / C / D / E / F / M, and it was determined by HPLC whether they produced transglucosyl Stevioside, transglucosylated rubusoside, transglucosylated dulcoside A, and transglucosylated rebaudioside A / C / D / E / F / M. Specifically, based on Figures 1 to 9 In order to identify a substance newly produced by the reaction between the crude enzyme solution of the Lactobacillus malipus strain and the steviol glycosid...
Embodiment 3
[0051] Example 3: Effect of Temperature on the Synthesis of Transglucosylated Steviol Glycosides
[0052] The effect of temperature was evaluated in the production of transglucosylated steviol glycosides by crude enzyme solutions derived from Lactobacillus malipus strains. Steviol glycosides (stevioside and rebaudioside A) and sugar were dissolved in 0.05M acetic acid buffer solution (pH 5.0), and a crude enzyme solution was added thereto, followed by heating at 10°C to 80°C React for 24 hours. After the reaction, the amount of the transglucosylated steviol glycoside in the reaction solution was analyzed by HPLC.
[0053] Figure 17 It is a graph showing the conversion rate of transglucosylated steviol glycosides (stevioside and rebaudioside A) according to temperature. based on Figure 17 , the conversion rate of the crude enzyme solution of the Lactobacillus malus strain to the transglucosylated steviol glycoside is as high as 10% to 70% at 10°C to 50°C.
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